Problems of isolating Borrelia burgdorferi from ticks collected in United Kingdom foci of Lyme disease

Livesley, M. A.; Carey, Dorothy; Gern, Lise; Nuttall, Patricia A.

doi:10.1111/j.1365-2915.1994.tb00159.x

Cited by 19 publications

(7 citation statements)

References 17 publications

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“…Our results suggest that isolates may be selected against even more rapidly, perhaps in the initial culture. B. burgdorferi could be cultured in BSK II medium from only 7 to 10% of Ixodes ricinus ticks in the United Kingdom that were PCR positive with ospA primers (25). Likewise, B. burgdorferi sensu lato was cultured in BSK II medium from only 23% of I. ricinus ticks from Valais, Switzerland, that were positive by indirect immunofluorescence detection (34).…”

Section: Discussionmentioning

confidence: 99%

Culturing selects for specific genotypes of Borrelia burgdorferi in an enzootic cycle in Colorado

et al. 1997

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In Colorado, Borrelia burgdorferi sensu stricto, the etiologic agent of Lyme disease, is maintained in an enzootic cycle between Ixodes spinipalpis ticks and Neotoma mexicana rats (27). The frequencies of flagellin (fla), 66-kDa protein (p66), and outer surface protein A (ospA) alleles were examined in 71 B. burgdorferi isolates from samples from Colorado. Approximately two-thirds of these samples were isolates from I. spinipalpis ticks that had been cultured in BSK-H medium prior to DNA extraction. The remaining samples were from total DNA extracted directly from infected I. spinipalpis ticks. A portion of each gene was amplified by PCR and screened for genetic variability by single-strand conformation polymorphism (SSCP) analysis. We identified three alleles in the fla gene, seven in the p66 gene, and seven in the ospA gene. Sequencing verified that the amplified products originated from B. burgdorferi template DNA and indicated 100% sensitivity and specificity of the SSCP analysis. The frequencies of the p66 and ospA alleles were significantly different between cultured and uncultured spirochetes. The number of three-locus genotypes and the genetic diversity of alleles at all loci were consistently lower in cultured spirochetes, suggesting that culturing of B. burgdorferi in BSK-H medium may select for specific genotypes.

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Section: Discussionmentioning

confidence: 99%

Culturing selects for specific genotypes of Borrelia burgdorferi in an enzootic cycle in Colorado

et al. 1997

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“…Polymerase Chain Reaction. DNA from tick and biopsy samples was extracted using phenol-chloroform (Livesley et al 1994. B. burgdorferi s.l.…”

Section: Methodsmentioning

confidence: 99%

Competence of Pheasants as Reservoirs for Lyme Disease Spirochetes

Kurtenbach

Carey

Hoodless

et al. 1998

Journal of Medical Entomology

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Pheasants, Phasianus colchicus L., constitute a major part of the ground-feeding avifauna of England and Wales and are important hosts to immature stages of Ixodes ricinus L., the principal tick vector of Lyme borreliosis spirochetes in Europe. Therefore, their competence as hosts for Borrelia burgdorferi Johnson, Schmid, Steigerwalt & Brenner sensu lato was investigated. One group of pheasants was inoculated by needle with 1 x 10(6) cultured B. burgdorferi s.s. organisms, and a 2nd group of birds was infested with I. ricinus nymphs collected from a focus of Lyme borreliosis in southern England. Both bird groups were subjected to xenodiagnoses using uninfected I. ricinus nymphs. All recovered engorged ticks, as well as pheasant skin biopsies, were analyzed by a nested polymerase chain reaction targeting the 5S-23S rRNA genes of B. burgdorferi s.l. Both groups proved to be infective for ticks. The birds that were infected by tick bites proved to be significantly more infective for ticks (23% of the xenodiagnostic ticks positive) than those infected by needle (5%). The results show that pheasants can be infected experimentally with B. burgdorferi s.l., that they can pass the spirochetes to ticks and that their infectivity for ticks may persist as long as 3 mo. We conclude that pheasants are reservoir competent for Lyme borreliosis spirochetes and potentially play an important role in the maintenance of B. burgdorferi s.l. in England and Wales.

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“…Approaches based on the PCR (33), however, appear to be more accurate in assessing the diversity and distribution of borreliae in nature, as culturing may favor particular genotypes (27). Furthermore, strains of B. burgdorferi sensu lato prevalent in the United Kingdom (known to be variants of B. garinii, B. afzelii, B. burgdorferi sensu stricto, and B. valaisiana) have been found to be unusually difficult to isolate and culture from ticks and hosts by standard techniques (20). There is increasing evidence that the kinds of borreliae in ticks and hosts vary considerably (9,14,28,29,32,33).…”

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Differential Transmission of the Genospecies of Borrelia burgdorferi Sensu Lato by Game Birds and Small Rodents in England

Kurtenbach

Peacey

Rijpkema³

et al. 1998

Appl Environ Microbiol

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The genetic diversity of Borrelia burgdorferi sensu lato was assessed in a focus of Lyme borreliosis in southern Britain dominated by game birds. Ticks, rodents, and pheasants were analyzed for spirochete infections by PCR targeting the 23S-5S rRNA genes, followed by genotyping by the reverse line blot method. In questingIxodes ricinus ticks, three genospecies of B. burgdorferi sensu lato were detected, with the highest prevalences found for Borrelia garinii and Borrelia valaisiana. B. burgdorferi sensu stricto was rare (<1%) in all tick stages. Borrelia afzelii was not detected in any of the samples. More than 50% of engorged nymphs collected from pheasants were infected with borreliae, mainly B. garinii and/orB. valaisiana. Although 19% of the rodents harboredB. burgdorferi sensu stricto and/or B. gariniiin internal organs, only B. burgdorferi sensu stricto was transmitted to xenodiagnostic tick larvae (it was transmitted to 1% of the larvae). The data indicate that different genospecies of B. burgdorferi sensu lato can be maintained in nature by distinct transmission cycles involving the same vector tick species but different vertebrate host species. Wildlife management may have an influence on the relative risk of different clinical forms of Lyme borreliosis.

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Problems of isolating Borrelia burgdorferi from ticks collected in United Kingdom foci of Lyme disease

Cited by 19 publications

References 17 publications

Culturing selects for specific genotypes of Borrelia burgdorferi in an enzootic cycle in Colorado

Culturing selects for specific genotypes of Borrelia burgdorferi in an enzootic cycle in Colorado

Competence of Pheasants as Reservoirs for Lyme Disease Spirochetes

Differential Transmission of the Genospecies of Borrelia burgdorferi Sensu Lato by Game Birds and Small Rodents in England

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